Magnetic quaternary ammonium polymer bearing porous agarose for selective extraction of Aristolochic acids in the plasma.

Aristolochic acids (AAs) naturally occurring in the herbal genus Aristolochia are associated with a high risk of kidney failure, multiple tumors and cancers. However, approaches with high selectivity and rapidity for measuring AAs in biological samples are still inadequate. Inspired by the mechanism of AAs-induced nephrotoxicity, we designed a hybrid magnetic polymer-porous agarose (denoted as MNs@SiO2M@DNV-A), mimicking the effect of basic and aromatic residues of organic anion transporter 1 (OAT1) for efficient enriching aristolochic acid I (AA I) and aristolochic acid II (AA II) in the plasma. The monomers of vinylbenzyl trimethylammonium chloride (VBTAC), N-vinyl-2-pyrrolidinone (NVP) and divinylbenzene (DVB) were employed to construct the polymer layer, which provided a selective adsorption for AAs by multiple interactions. The porous agarose shell contributed to remove interfering proteins in the plasma samples. A magnetic solid-phase extraction (MSPE) based on the proposed composite enhanced the selectivity toward AA I and AA II in the plasma samples. In combination of HPLC analysis, the proposed method was proved to be applicable to fast and specific quantification of AAs in blood samples, which was characterized by a good linearity, high sensitivity, acceptable recovery, excellent repeatability and satisfactory reusability.

Metal-organic framework grafted with melamine for the selective recognition and miniaturized solid phase extraction of aristolochic acid â from traditional Chinese medicine.

Aristolochic acid Ⅰ is a nephrotoxic compound and exist in some traditional Chinese medicines at trace level. Up to now, specific enrichment of aristolochic acid Ⅰ remains important procedure and key problem in its analysis. In this study, melamine was proposed as the recognition unit and grafted on the surface of metal-organic framework to fabricate a specific material for aristolochic acid Ⅰ. This material was prepared by using a two-step strategy and the preparation process was optimized. The physical and chemical properties were investigated using scanning electron microscopy, Fourier-transfer infrared spectroscopy, X-ray diffraction and nitrogen adsorption-desorption techniques. Adsorption properties were evaluated by binding experiments. The melamine modified material exhibited a uniform morphology, high specific surface area (460.20 m2 g-1), high adsorption capacity (25.57 mg g-1), fast mass transfer rate and excellent selectivity. Further, a specific and sensitive method was established by using this material as adsorbent of mini-solid phase extraction. The limit of detection was as low as 0.02 µg mL-1. Therefore, melamine modified metal-organic framework is an ideal adsorbent for the recognition and enrichment of aristolochic acid Ⅰ.

Aristololactam alkaloids from the roots of Dasymaschalon rostratum.

A novel aristololactam alkaloid, dasymalactam A (1), together with nine known analogues (2-10), were isolated from the roots of Dasymaschalon rostratum. Their structures were elucidated by IR, NMR and MS spectrums and comparisons with data reported in the literature. All compounds demonstrated weak cytotoxicity against Hela, MCF-7, A-549, MGC-803, and COLO-205 human cancer cell lines.

Chemotaxonomy of the ethnic antidote Aristolochia indica for aristolochic acid content: Implications of anti-phospholipase activity and genotoxicity study.

ETHNOPHARMACOLOGICAL RELEVANCE: Aristolochia indica L. (Aristolochiaceae) is a common medicinal plant described in many traditional medicine as well as in Ayurveda used against snakebites. Besides, the plant has also been reported traditionally against fever, rheumatic arthritis, madness, liver ailments, dyspepsia, oedema, leishmaniasis, leprosy, dysmenorrhoea, sexual diseases etc. The plant is known to contain its major bioactive constituent aristolochic acid (AA) known for its anti-snake venom, abortifacient, antimicrobial and antioxidant properties. MATERIALS AND METHODS: This present work describes a validated, fast and reproducible high performance thin layer chromatography (HPTLC) method to estimate AA from the roots of 20 chemotypes of A. indica procured from 20 diverse geographical locations from the state of West Bengal, India. Further, an evidence-based approach was adopted to investigate the reported anti-venom activity of the aqueous extracts of the A. indica roots by assessing its phospholipase A2 (PLA2) inhibitory properties since PLA2 is a major component of many snake-venoms. Finally, the cytotoxicity and genotoxicity of the aqueous root extract of the Purulia (AI 1) chemotype were assessed at various concentrations using Allium cepa root meristematic cells. RESULTS: The highest amount of AA (7643.67 µg/g) was determined in the roots of A. indica chemotype collected from Purulia district followed by the chemotypes collected from Murshidabad, Jalpaiguri and Birbhum districts (7398.34, 7345.09 and 6809.97 µg/g respectively). This study not only determines AA in the plants to select pharmacologically elite chemotypes of A. indica, but it also identifies high AA producing A. indica for further domestication and propagation of the plants for pharmacological and industrial applications. The method was validated via analyzing inter-day and intra-day precision, repeatability, reproducibility, instrumental precision, limit of detection (LOD) and limit of quantification (LOQ) and specificity. Chemotypes with high AA content exhibited superior anti-PLA2 activity by selectively inhibiting human-group PLA2. Moreover, A. indica root extract significantly inhibited mitosis in Allium cepa root tips as a potent clastogen. CONCLUSIONS: The present quick, reproducible and validated HPTLC method provides an easy tool to determine AA in natural A. indica plant populations as well as in food and dietary supplements, a potential antivenin at one hand and a possible cause of aristolochic acid nephropathy (AAN) at another. Besides, the cytotoxic and mitotoxic properties of the root extracts should be used with caution especially for oral administration.

Adenine-coated magnetic multiwalled carbon nanotubes for the selective extraction of aristolochic acids based on multiple interactions.

A method is described for the functionalization of magnetic carbon nanotubes to recognize aristolochic acid Ⅰ and Ⅱ. 3-Glycidyloxypropyltrimethoxysilane was used as a coupling agent to immobilize adenine on a solid support. The morphology and structure of adenine-coated magnetic carbon nanotubes was investigated using transmission electron microscopy (TEM), scanning electron microscopy (SEM), X-ray diffraction (XRD) and a vibrating sample magnetometer (VSM). The adsorption performance of the adenine-coated magnetic carbon nanotubes was evaluated via adsorption isotherms, the kinetics and selectivity tests. The adsorption capacity of the adenine-functionalized sorbent for aristolochic acid Ⅰ was determined to be 24.5 µg mg-1. By combining magnetic solid phase extraction with HPLC detection, a method was developed to enrich and detect aristolochic acids used in traditional Chinese medicine. A satisfactory recovery (92.7 - 97.5% for aristolochic acid Ⅰ and 92.6 - 99.4% for aristolochic acid Ⅱ) and an acceptable relative standard deviation (<4.0%) were obtained.

Genome-wide transcriptional analysis of Aristolochia manshuriensis induced gastric carcinoma.

Context: Aristolochia manshuriensis Kom (Aristolochiaceae) (AMK) is known for toxicity and mutagenicity.Objective: The tumorigenic role of AMK has yet to be understood.Materials and methods: AMK extracts were extracted from root crude drug. SD (Sprague Dawley) rats underwent gavage with AMK (0.92 g/kg) every other day for 10 (AMK-10) or 20 (AMK-20) weeks. Stomach samples were gathered for histopathological evaluation, microarray and mRNA analysis.Results: The gastric weight to body weight ratio (GW/BW) is 1.7 in the AMK-10 cohort, and 1.8 in AMK-20 cohort compared to control (CTL) cohort. Liver function was damaged in AMK-10 and AMK-20 rats compared to CTL rats. There were no significant changes of CRE (creatinine) in AMK-10 and AMK-20 rats. Histopathological analysis revealed that rats developed dysplasia in the forestomach in AMK-10 rats, and became gastric carcinoma in AMK-20 rats. Genes including Mapk13, Nme1, Gsta4, Gstm1, Jun, Mgst2, Ggt6, Gpx2, Gpx8, Calml3, Rasgrp2, Cd44, Gsr, Dgkb, Rras, and Amt were found to be critical in AMK-10 and AMK-20 rats. Pik3cb, Plcb3, Tp53, Hras, Myc, Src, Akt1, Gnai3, and Fgfr3 worked in AMK-10 rats, and PDE2a and PDE3a played a pivotal role in AMK-20 rats.Discussion and conclusions: AMK induced benign or malignant gastric tumours depends on the period of AMK administration. Genes including Mapk13, Nme1, Gsta4, Gstm1, Jun, Mgst2, Ggt6, Gpx2, Gpx8, Calml3, Rasgrp2, Cd44, Gsr, Dgkb, Rras, Amt, Pik3cb, Plcb3, Tp53, Hras, Myc, Src, Akt1, Gnai3, Fgfr3, PDE2a, and PDE3a were found to be critical in aristolochic acid-induced gastric tumour process.

Isolation and purification of six aristolochic acids with similar structures from Aristolochia manshuriensis Kom stems by pH-zone-refining counter-current chromatography.

Aristolochic acids (AAs), the major components in Aristolochia manshuriensis Kom stems (AMK), may cause Chinese herb nephropathy during clinical application. Therefore, it is necessary to distinguish AMK from other herbs and Chinese medicines using AAs with high purity as standards. So, an efficient method for separation and purification of AAs is required because of their similar structures. In this study, six AAs with purities of >98% were obtained by pH-zone-refining counter-current chromatography (PZRCCC) in a single run. The optimum two-phase solvent system was petroleum ether-ethyl acetate-methanol-water (3:7:3:7, v/v). Triethylamine (10 mM) was added to the aqueous mobile phase and trifluoroacetic acid (10 mM) to the organic stationary phase. As a result, 9.7 mg aristolochic acid IIIa, 12.0 mg aristolochic acid IVa, 32.2 mg aristolochic acid II, 103.7 mg aristolochic acid I, 24.6 mg aristolic acid II, and 26.1 mg aristolic acid I were obtained from 800 mg AAs crude extract. The elution order of AAs during PZRCCC separation corresponded with the pKa values and hydrophobicities of the target compounds. PZRCCC is an efficient method for isolation of AAs with similar structures.

Electromembrane extraction of aristolochic acids: New insights in separation of bioactive ingredients of traditional Chinese medicines.

Aristolochic acid (AA) I and AA II, which have been classified as carcinogenic to human and have been proven to be nephrotoxic, are bioactive ingredients of many traditional Chinese medicines (TCMs). Thus, development of an efficient approach for separation and determination of AA I and AA II in biological samples and herbal plants is of significance. Herein, electromembrane extraction (EME) was for the first time used to separate AA I and AA II. It is noted that also for the first time 1-decanol was discovered and used as an efficient SLM solvent for EME of acidic compounds. The proposed EME system was used to extract AA I and AA II from urine samples (recovery≥68%). The approach of EME combined with LC-MS (EME-LC/MS) was evaluated using urine samples. The linear range for AA I and AA II was 10-1000 ng mL-1 (R2≥0.9970), and the limits of detection (LOD, S/N = 3) for AA I and AA II were 2.7 and 2.9 ng mL-1, respectively. Finally, this EME-LC/MS approach was employed to discover AA I and AA II in the herbal plants. In addition, using standard addition method, AA I in Aristolochicaceae-Liao Asarum (ALA) and Radix Aristolochice (RA) were 0.23 and 2044.13 µg g-1, and AA II in ALA and RA were 0 and 338.48 µg g-1, respectively. The repeatability of EME-LC/MS at all cases for both urine samples and herbal plants was below 15% (RSD-value). We believe that EME would be a useful tool to isolate bioactive ingredients of TCMs from complex samples for different purposes.

Isolation of aristolochic acid I from herbal plant using molecular imprinted polymer composited ionic liquid-based zeolitic imidazolate framework-67.

Aristolochic acid I is a toxic compound found in the genus of Aristolochia plants, which are commonly used as herbal cough treatment medicines. To remove the aristolochic acid I in extract efficiently and selectively, a molecularly imprinted polymer composed of ethylimidazole ionic liquid-based zeolitic imidazolate framework-67 was synthesized and used as the adsorbent. Under the conditions optimized by the software design expert, the sorbent showed highest adsorption amount of 34.25 mg/g in methanol/water (95:5, v/v) at 39°C for 138 min. The sorbent was then applied to solid phase extraction to isolate aristolochic acid I from the extract of the herbal plant Fibraurea Recisa Pierre. 0.043 mg/g of aristolochic acid I was obtained after the loading, washing, and elution processes. The limit of detection of 2.41 × 10-5  mg/mL and good recoveries provided evidence for the accuracy of this method.

Preparation of magnetic molecularly imprinted polymers functionalized carbon nanotubes for highly selective removal of aristolochic acid.

In this work, we presented the preparation of magnetic carbon nanotubes (MCNTs) functionalized with molecularly imprinted polymers (MIPs) for effective removal of aristolochic acid I (AAI) in traditional Chinese medicine (TCM). MCNTs@AAI-MIPs was obtained via a facile and environmental friendly sol-gel process. Firstly, MCNTs were synthesized by a solvothermal method. Then, the template molecules were self-assembled with the functional monomer phenyltrimethoxysilane (PTMOS) in the presence of ethanol and water. Finally, AAI-MIPs film was coated on the MCNTs to obtain product MCNTs@AAI-MIPs using tetraethyl-orthosilicate (TEOS) as cross-linker. The morphology and structure of prepared MIPs were characterized by Fourier transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), X-ray diffraction (XRD), nitrogen gas adsorption and vibrating sample magnetometer (VSM). The adsorption properties were demonstrated by kinetic, isothermal and selective adsorption experiments. The results showed that the imprinted nanocomposites exhibited fast separation rate (10 s), high adsorption capacity (18.54 µg∙mg-1), short kinetic equilibrium time (15 min), and good selectivity to template molecule with imprinting factor (IF) of 3.17. A regression equation (y=57294x-4734.1) with good linearity was obtained in the concentration range of 0.1-200 µg∙mg-1 for AAI with a correlation coefficient (R2) of 0.9998. The limit of detection (LOD, S/N=3) was 0.034 µg∙mg-1. Moreover, high recoveries ranged from 80% to 110% (RSD=3.27%-8.16%) were received in spiked TCM samples. The results suggested that the proposed MCNTs@AAI-MIPs could efficiently and specifically capture AAI from an actual complex TCM samples.

Combined magnetic porous molecularly imprinted polymers and deep eutectic solvents for efficient and selective extraction of aristolochic acid I and II from rat urine.

Aristolochia and related plants contain nephrotoxins and mutagens in the form of aristolochic acids (AAs). However, there is still lack of a fast and specific method for monitoring AAs in biological samples. Herein, we synthesized a hybrid magnetic mesoporous carbon-molecularly imprinted polymers (MMC@MIPs) as a novel magnetic solid-phase extraction (MSPE) adsorbent for selective recognition of aristolochic acid I and II from rat urine samples. The choline chloride/glycol-based deep eutectic solvent (DES) and indomethacin were used as the eluent and dummy template molecule accordingly. The morphology, structure property and surface groups of the prepared materials were investigated in sequence, and the optimum conditions of the MMC@MIPs-MSPE procedure were also optimized well. Results showed that the proposed method had a relatively satisfactory recovery (86.7-94.3%), with low standard deviation (<4.85%) and acceptable correlation coefficients (0.991-0.996). Overall, this work not only provides an inexpensive and eco-friendly method to fabricate MMC@MIPs, but also develops a highly promising approach for the detection of aristolochic acid I and II in biological samples.

Dual ionic liquid-immobilized silicas for multi-phase extraction of aristolochic acid from plants and herbal medicines.

A convenient, and efficient multi-phase extraction method was used to isolate aristolochic acid from real samples. To increase the efficiency, a new dual ionic liquid-immobilized silica was used as a sorbent and the effects of the adsorption conditions were investigated. IM-BIM@Sil in a methanol/water (60:40, v/v) solution at 60 °C extracted the highest amount of aristolochic acid (16.69 mg/g) compared to the other sorbents examined. The sorbent was then applied to the multi-phase extraction of aristolochic acid. After extraction eight times, 2.4-70.9 × 10-3 mg/g of aristolochic acid was isolated from nine real samples after several washing and elution steps. The recoveries of aristolochic acid ranged from 70.0% to 110.6% with relative standard deviations of 3.5%-9.1%, highlighting the accuracy of this method.

Hybrid-type carbon microcoil-chitosan composite for selective extraction of aristolochic acid I from Aristolochiaceae medicinal plants.

Aristolochic acid I is a nephrotoxic compound widely existing in many kinds of traditional Chinese medicines, especially in Aristolochiaceae medicinal plants. In this study, chitosan modified carbon microcoils were designed and prepared for the selective separation of aristolochic acid I from medicinal herbs. Successful modification of carbon microcoils was confirmed by scanning electron microscopy, Fourier-transfer infrared spectroscopy, elemental analysis, X-ray photoelectron spectroscopy, and thermogravimetric analyses. The effects of adsorption conditions were investigated and it was determined that the adsorption of aristolochic acid I was controlled by pH. Adsorption isotherms, kinetics, and selectivity tests were performed to evaluate the adsorption capacity and selectivity of the modified carbon microcoils. The chitosan modified carbon microcoils exhibited excellent binding ability (77.72 mg g-1) and satisfactory selectivity. Finally, this material was used in solid phase extraction combined with HPLC to enrich and detect aristolochic acid I from medicinal plants. The detector response for aristolochic acid I was linear from 0.5 to 150 mg L-1, and the recoveries of aristolochic acid I ranged from 73.61 to 77.73% with the relative standard deviations of less than 5%. Thus, chitosan modified carbon microcoils were ideal adsorbents for the selective extraction of aristolochic acid I from Aristolochiaceae plants.

Fermentation of Aristolochia debilis by six different medicinal fungi and identification of aristolochic acid derivates in their products by HPLC-ESI-TOF-MS.

To analyze the content change of the nephrotoxic substances, aristolochic acid derivates (AAs) in the roots of Aristolochia debilis and the products generated from the solid-state fermentation of six different medicinal fungi by HPLC-ESI-TOF-MS, the chromatographic separation was carried out on C18 column at 30°C with the DAD detector. The elution was performed using the mobile phase of acetonitrile (A) and 0.2% acetic acid (B). Several new peaks were found in the scale of 0-20 min elution of HPLC diagram in the fermentation products. The ESI-MS detection (negative ion mode) was carried out by post-column flow splitting method following the automatic injection. Seven AAs in the fermentation products and A. debilis were deduced, which were recognized as AAIa or IIIa (1), AAVIa (2), AAIVa, Va, VIIa or VIIIa (3); AAII (4); AAIII (5); AAI (6); AAIV or VII (7). The areas of almost all these seven components existing originally in the corresponding crude drug decreased after the fermentation process, suggesting that fermentation is an effective way of lowering the nephrotoxicity induced by AAs in Chinese medicines similar with A. debilis. In addition, Optimized HPLC-MS method is helpful to AAs content identification.

Natural Aristolochia Alkaloid Aristololactam-ß-D-glucoside: Interaction with Biomacromolecules and Correlation to the Biological Perspectives.

BACKGROUND: Natural aristolochia alkaloids have attracted the attention of both chemists and biologists from the stand point of their structural and pharmacological aspects. Many of the compounds isolated in this group are potent tumor inhibitors. These are divided into nitrophenanthrinic acid, phenanthrene lactams and isoquinoline alkaloids. A number of structure-activity studies have been performed on aristolochia alkaloids. Of particular interest is the molecule with the ß-D-glucoside moiety that has similarity to the clinical anticancer agent daunomycin. OBJECTIVE: The anticancer activity of aristololactam-ß-D-glucoside has been thought to be due to its DNA and RNA binding activities among other actions. In this article we focus on the physicochemical property of this alkaloid and the structural and functional aspects of its binding to different nucleic acid and protein structures. METHODS: This review highlights a large number of biophysical studies employing various analytical techniques like absorbance, fluorescence, circular dichroism, thermal melting, viscosity, IR study, isothermal calorimetry and differential scanning calorimetry. RESULT: The detailed binding mechanism in terms of the structural and thermodynamic aspects at the molecular level has been discussed. CONCLUSION: This review enables to assess the high potential of developing aristololactam-ß-Dglucoside and related alkaloids as therapeutic agents.

Aristololactam derivatives from the fruits of Aristolochia contorta Bunge.

Three new aristololactam derivatives, aristololactam W-Y (1-3), and three known compounds (4-6) were isolated from the fruits of Aristolochia contorta Bunge. Compounds 1 and 2 represent the first example of an N-CH2OCH3 aristololactam derivative from natural products. Their structures were elucidated by 1D/2D NMR and HRESIMS spectra. All of the isolated compounds were evaluated for their insecticidal activity against 4th instar larvae of Aedes aegypti. Compound 4 displayed insecticidal activity with LC50 value of 3.54 µg/mL.

Aristolic Acid Derivatives from the Bark of Antidesma ghaesembilla.

Antidesma ghaesembilla is an important medicinal and food plant in many Asian countries. Ten substances could be isolated from the dichloromethane and methanol extract: sitostenone (3), daucosterol (4), chavibetol (5), asperphenamate (6), protocatechuic acid (7), vanillic acid-4-O-ß-D-glucoside (8), 1-O-ß-D-glucopyranosyl-3-O-methyl-phloroglucinol (9), and aristolic acid II-8-O-ß-D-glucoside (10), and two new aristolic acid derivatives, 10-amino-5,7-dimethoxy-aristolic acid II (= 6-amino-9,11-dimethoxyphenanthro[3,4-d]-1,3-dioxole-5-carboxylic acid; 1) and 5,7-dimethoxy-aristolochic acid II (= 9,11-dimethoxy-6-nitrophenantro[3,4-d]-1,3-dioxole-5-carboxylic acid; 2). Exposure to humans of some of these compounds is associated with a severe disease today known as aristolochic acid nephropathy. Therefore, the traditional usage of this plant has to be reconsidered carefully.

Aristolochic acid derivatives from the rhizome of Arisolochia championii.

Four new aristolochic acid derivatives aristchamic A (1), aristchamic B (2), aristochamic C (3a), aristchamic D (3b) and one new aristolactam aristolactam-CV (4), together with 10 known compounds (5-14), were isolated from the rhizomes of Aristolochia championii. Their structures were assigned by detailed analysis of MS and NMR spectroscopic data. All of the isolated compounds were evaluated for their cytotoxic activities against HCT-116, HepG2, BGC-823, NCI-H1650, and A2780 cell. Compound 1 exhibited significant cytotoxic activity against HCT-116, HepG2, BGC-823, and NCI-H1650, with IC50 values of 0.50, 7.37, 2.66, and 0.75µM, respectively.

New aristolochic acid and other chemical constituents of Aristolochia maurorum growing wild in Jordan.

Investigation of the chemical constituents of Aristolochia maurorum growing wild in Jordan resulted in the isolation and characterisation of one new compound in addition to 19 known compounds. The new compound was identified as aristolochic acid II alanine amide (14). The other known compounds were the following: palmitic acid (1), ß-sitosterol (2), E-ethyl-p-coumarate (3), Z-ethyl-p-coumarate (4), aristolochic acid IV methyl ester (5), aristolactam I (6), loliolide (7), (+)-dehydrovomifoliol (8), glycerol-1-palmitate (9), aristolochic acid I (10), E-p-coumaric acid (11), E-N-coumaroyltyramine (12), ß-sitosteryl glucoside (13), aristolochic acid IV (15), aristolochic acid III (16), esculetin (17), uracil (18), shepherdine (19) and adenosine (20). The isolated compounds were characterised by different spectroscopic methods including NMR (1D and 2D), UV, IR and HRESIMS.

Preparation and adsorption properties of molecularly imprinted polymer via RAFT precipitation polymerization for selective removal of aristolochic acid I.

The molecularly imprinted polymers (MIP) were prepared via aqueous RAFT precipitation polymerization, with aristolochic acid I (AAI) as the template molecule, AA as the functional monomer, EGDMA as the cross-linker, AIBN as the chain initiator, CTP as the chain transfer agent and 80% (g/g) DMF-aqueous solution as the porogen. The differential UV-vis spectra revealed that a cooperative hydrogen-bonding complex between AAI and AA might be formed at the molar ratio of 1:3 in prepolymerized system. The synthesized MIPs were characterized by FTIR spectra, solid UV-visible absorption spectra, nitrogen adsorption-desorption isotherms and scanning electron microscope, which proved that the MIPs and NIPs have the similar chemical structures and the adding of AAI could affect the size and morphology of the microspheres. UV-visible absorption spectra and High-performance liquid chromatography (HPLC) was used to investigate the adsorption and recognition properties of the MIPs. The Scatchard isotherm model described that the binding sites independently acted. The Langmuir isotherm model suggested an excellent imprinting effect owing to the presence of a large number of specific binding sites on the MIP. The Freundlich model indicated that the AAI could be readily absorbed by MIP. Selective absorption of the template molecule was demonstrated in presence of its analogous compounds, benzoic acid and nitrobenzene. The recycling experiments implied that the MIP could be reused to further selective recognition and separation to AAI for six times at least. MIP was developed for removal of AAI from the Aristolochia manshuriensis extraction. The results indicated that 25mg of MIP could remove the AAI below the HPLC detection limits (6.47ngmL-1) from 5.0mL of the extraction (CAAI=0.0018mgmL-1) with the recovery of AAI to 91.50% (n=3, SD=4.24%). Therefore, it is clearly revealed that the MIP can be a useful tool to remove toxic compounds from natural products.